old male balb c mice  (Jackson Laboratory)

Journal: Cell Reports Medicine

Article Title: A dual-adjuvanted HA stem nanoparticle vaccine elicits a multifunctional antibody response that is associated with protection in newborn monkeys

doi: 10.1016/j.xcrm.2026.102746

Figure Lengend Snippet: Conjugation of R848 to H1ssF-Cys nanoparticles (A) Representative image of H1ssF-Cys mutant. (B and C) Size (B) and PDI (C) analysis of H1ssF (WT and Cys mutant) using DLS. (D) Schematic for conjugation of R848 to H1ssF-Cys. BALB/c mice ( n = 3–12 across 3–4 experiments depending on the vaccine tested) were vaccinated and NC99 stem-specific Abs were quantified 14 days p.v. (E) BALB/c mice ( n = 3–12 across 3–4 experiments depending on the vaccine tested) were vaccinated and NC99 stem-specific Abs were quantified 14 days p.v. The dotted line indicates the limit of detection for the assay. TT was defined as the highest dilution with an OD 450 greater than three times the assay background. Lines represent the mean ± SEM. (F) Negatively stained TEM images of the conjugated H1ssF-R848. The scale bar lengths are as follows: 50 nm (A and F, left), 10 nm (F, right). Not significant p ≥ 0.05 (not indicated on graph), ∗∗ p ≤ 0.01, ∗∗∗∗ p ≤ 0.0001.

Article Snippet: Six-week-old female BALB/c , Charles River Laboratories , Cat# 028.

Techniques: Conjugation Assay, Mutagenesis, Staining

Journal: Cell Reports Medicine

Article Title: Reprogramming T cell-myeloid crosstalk overcomes immune resistance in colorectal cancer

doi: 10.1016/j.xcrm.2026.102786

Figure Lengend Snippet: MMRd tumors responding to anti-PD-1 are highly infiltrated by MHC+ C1Q+ macrophages (A) A total of 200,000 CT26 tumor cells, either deleted or not for MSH2, were allowed to grow subcutaneously for 28 or 35 days in BALB/c mice, with or without anti-PD-1 administered twice a week after 14 days ( N = 10 replicates, Mann-Whitney U test, p value <0.05). Validation with 4 other orthotopic models was performed in . (B and C) Quantification of differential lymphocyte infiltration expression according to the MSI status and ICB through spectral flow cytometry at day 28 ( N = 6 replicates, Mann-Whitney U test, p value <0.05). Only significant results are shown. (D) scRNA-seq UMAP of immune cells in CT26 WT and MSH2 KO tumors with or without anti-PD-1 therapy ( N = 3 replicates per condition). (E) UMAP of immune clusters subsequent to Leiden clustering by scanpy. Percentage of cells in each immune cluster. (F) Gene expression within each immune cluster. (G–J) We reanalyzed human MMRd CRC patient data previously published, focusing on patients who responded or did not respond to anti-PD-1 therapy alone, and the underlying T cell and macrophage profiles. ( N = 9 patients). (G) scRNA-seq UMAP of immune cells in patients who responded (pathological complete response, pCR) or did not respond (non-pCR) to anti-PD-1 therapy. (H) UMAP of immune clusters subsequent to Leiden clustering by scanpy. (I) Percentage of cells in each immune cluster. (J) Gene expression within each immune cluster.

Article Snippet: 2 ∗ 10ˆ5 WT CT26 cells or CT26 MSH2 KO or 4T1 MSH2 KO or B16F10 MSH2 KO or MC38 cells in 100 μL of PBS were orthotopically or subcutaneously injected in 6-week old BALB/c or BL6 mice (Jackson Laboratories) (IACCR202500000183/IACUC-2029-0062).

Techniques: MANN-WHITNEY, Biomarker Discovery, Expressing, Flow Cytometry, Gene Expression

Journal: Cell Reports Medicine

Article Title: Reprogramming T cell-myeloid crosstalk overcomes immune resistance in colorectal cancer

doi: 10.1016/j.xcrm.2026.102786

Figure Lengend Snippet: A high diversity of CCL5+ PRF1+ T cell clones, DCs, and MHC+ C1Q+ macrophages colocalize in MMRd tumors responding to anti-PD-1 A total of 200,000 CT26 tumor cells, either deleted or not for MSH2, were allowed to grow for 28 or 35 days in BALB/c mice, with or without anti-PD-1 administered twice a week after 14 days ( N = 10 replicates, Mann-Whitney U Test, p value <0.05). (A) TCRseq paired with scRNAseq UMAP of immune cells in CT26 WT and MSH2 KO tumors with or without anti-PD-1 therapy ( N = 3 replicates per condition). UMAP of immune clusters subsequent to Leiden clustering by scanpy and clonal expansion of T cell clones. Each dot is a T cell. (B) Identification of T cell clones, where each clone is uniquely numbered, and their frequency is demonstrated as a pie chart. The number of cells in each clonotype is indicated by the size of each bubble. Selected gene expression within each main T cell clone is shown at the bottom (clone of interest: blue vs. other clones: orange). (C) Normalized Shannon entropy according to ICB status. (D) scRNA-seq data and overexpressed genes in T cells, macrophages, and DCs (cluster of interest: blue vs. others clusters: orange). (E) Spatial localization of each single-cell immune cluster in MSH2 KO tumors with anti-PD-1 therapy. Neighborhood enrichment and quantification were performed using scanpy and squidpy .

Article Snippet: 2 ∗ 10ˆ5 WT CT26 cells or CT26 MSH2 KO or 4T1 MSH2 KO or B16F10 MSH2 KO or MC38 cells in 100 μL of PBS were orthotopically or subcutaneously injected in 6-week old BALB/c or BL6 mice (Jackson Laboratories) (IACCR202500000183/IACUC-2029-0062).

Techniques: Clone Assay, MANN-WHITNEY, Gene Expression, Single Cell

Journal: Cell Reports Medicine

Article Title: Reprogramming T cell-myeloid crosstalk overcomes immune resistance in colorectal cancer

doi: 10.1016/j.xcrm.2026.102786

Figure Lengend Snippet: TCF+ T cells, CD4 + and CD8 + T cells, neutrophils, and MHC+ macrophages orchestrate the response to targeted checkpoint/myeloid combinations CT26 tumors deleted for MSH2 are growing for 28 days in BALB/c mice. (A) scRNA-seq UMAP of immune cells in CT26 MSH2 KO tumors with or without anti-PD-1/CTLA4/LAG3/TREM2 therapy. UMAP of immune clusters subsequent to Leiden clustering by scanpy. Percentage of cells in each immune cluster and gene expression within each immune cluster ( N = 3 replicates per condition). (B) Spatial RNA-seq localization of each single-cell immune cluster in MSH2 KO tumors following ICB. Neighborhood enrichment and quantification were performed using scanpy and squidpy . (C) Quantification by flow cytometry of differential immune checkpoint and lymphocyte infiltration expression according to response to PD-1/LAG3/TREM2 blockade. N = 8 replicates, t test, p value < 0.05. Data are represented as mean ± SEM. Mice with a decrease in tumor volume following therapy are considered as responders, while mice with an increase in tumor volume following therapy are considered as non-responders. (D) Immune composition in spleens and lymph nodes was also measured (flow cytometry). N = 8. Mann-Whitney U test, p value < 0.05. Data are represented as mean ± SEM. (E) Ten thousand MMRd CRC patient-derived cells are growing for 3 days as spheroids and 100,000 peripheral blood mononuclear cells (PBMCs) from 3 donors are activated with IL-15. Then, spheroids and PBMCs are incubated for 3 days. PBMCs were incubated before with anti-PD-1, anti-TIM3, and anti-LAG3 (10 μg/mL). Immune cells are previously labeled by CellTracker (red). Spheroid cell nuclei are labeled with NucBlue (blue). Spheroids were washed to remove non-infiltrating immune cells before analysis. 3D bi-photon imaging and flow cytometry staining of spheroids after dissociation. Quantification of differential immune checkpoint expression and lymphocyte infiltration according to ICB. N = 4. Mann-Whitney U test, p value < 0.05. Data are represented as mean ± SEM.

Article Snippet: 2 ∗ 10ˆ5 WT CT26 cells or CT26 MSH2 KO or 4T1 MSH2 KO or B16F10 MSH2 KO or MC38 cells in 100 μL of PBS were orthotopically or subcutaneously injected in 6-week old BALB/c or BL6 mice (Jackson Laboratories) (IACCR202500000183/IACUC-2029-0062).

Techniques: Gene Expression, RNA Sequencing, Single Cell, Flow Cytometry, Expressing, MANN-WHITNEY, Derivative Assay, Incubation, Labeling, Imaging, Staining